Effect of bleaching on sound enamel and with early artificial caries lesions using confocal laser microscopy.

The aim of this study was to evaluate effect of bleaching agents on sound enamel (SE) and enamel with early artificial caries lesions (CL) using confocal laser scanning microscopy (CLSM). Eighty blocks (4 x 5 x 5 mm) of bovine enamel were used and half of them were submitted to a pH cycling model to induce CL. Eight experimental groups were obtained from the treatments and mineralization level of the enamel (SE or CL) (n=10). SE groups: G1 - unbleached (control); G2 - 4% hydrogen peroxide (4 HP); G3 - 4 HP containing 0.05% Ca (Ca); G4 - 7.5% hydrogen peroxide (7.5 HP) containing amorphous calcium phosphate (ACP). CL groups: G5 - unbleached; G6 - 4 HP; G7 - 4 HP containing Ca; G8 - 7.5 HP ACP. G2, G3, G6, G7 were treated with the bleaching agents for 8 h/day during 14 days, while G4 and G8 were exposed to the bleaching agents for 30 min twice a day during 14 days. The enamel blocks were stained with 0.1 mM rhodamine B solution and the demineralization was quantified using fluorescence intensity detected by CLSM. Data were analyzed using ANOVA and Fisher's tests (α=0.05). For the SE groups, the bleaching treatments increased significantly the demineralization area when compared with the unbleached group. In the CL groups, no statistically significant difference was observed (p>0.05).The addition of ACP or Ca in the composition of the whitening products did not overcome the effects caused by bleaching treatments on SE and neither was able to promote remineralization of CL.

Effect of bleaching on sound enamel and with early artificial caries lesions using confocal laser microscopy

The aim of this study was to evaluate effect of bleaching agents on sound enamel (SE) and enamel with early artificial caries lesions (CL) using confocal laser scanning microscopy (CLSM). Eighty blocks (4 x 5 x 5 mm) of bovine enamel were used and half of them were submitted to a pH cycling model to induce CL. Eight experimental groups were obtained from the treatments and mineralization level of the enamel (SE or CL) (n=10). SE groups: G1 - unbleached (control); G2 - 4% hydrogen peroxide (4 HP); G3 - 4 HP containing 0.05% Ca (Ca); G4 - 7.5% hydrogen peroxide (7.5 HP) containing amorphous calcium phosphate (ACP). CL groups: G5 - unbleached; G6 - 4 HP; G7 - 4 HP containing Ca; G8 - 7.5 HP ACP. G2, G3, G6, G7 were treated with the bleaching agents for 8 h/day during 14 days, while G4 and G8 were exposed to the bleaching agents for 30 min twice a day during 14 days. The enamel blocks were stained with 0.1 mM rhodamine B solution and the demineralization was quantified using fluorescence intensity detected by CLSM. Data were analyzed using ANOVA and Fisher’s tests (α=0.05). For the SE groups, the bleaching treatments increased significantly the demineralization area when compared with the unbleached group. In the CL groups, no statistically significant difference was observed (p>0.05).The addition of ACP or Ca in the composition of the whitening products did not overcome the effects caused by bleaching treatments on SE and neither was able to promote remineralization of CL.

O objetivo deste estudo foi avaliar o efeito de agentes clareadores no esmalte sadio (ES) ou esmalte com lesão inicial de cárie artificial (LC), utilizando microscopia confocal laser de varredura (CLSM). Oitenta blocos (4 x 5 x 5 mm) de esmalte bovino foram usados, sendo que 40 destes foram desmineralizados com ciclagem de pH para induzir a LC. Oito grupos experimentais foram obtidos a partir dos tratamentos e condição do esmalte (ES ou LC), com n=10: Grupos ES: G1 - sem tratamento (controle); G2 - peróxido de hidrogênio 4% (PH4); G3: PH4 contendo 0,05% cálcio (Ca); G4 - peróxido de hidrogênio 7,5% (pH 7,5) contendo fosfato de cálcio amorfo (ACP). Grupos LC: G5 - não clareado; G6 - pH 4; G7 - pH 4 Ca; G8 - pH 7,5 ACP. Os grupos G2, G3, G6 e G7 foram tratados com o gel clareador por 8 h/dia durante 14 dias, enquanto as amostras dos grupos G4 e G8 foram submetidas ao agente clareador por 30 min/duas vezes ao dia, durante 14 dias. Os blocos de esmalte foram corados com solução de Rodamina B e a área fluorescente de desmineralização foi quantificada utilizando CLSM. Os dados foram submetidos a ANOVA e teste de Fisher (p<0,05). Para ES, os tratamentos clareadores aumentaram significativamente a área de desmineralização quando comparado com os grupos não clareados, entretanto, para LC não foi observado diferença estatística significante entre os grupos. A adição de ACP e Ca na composição dos géis clareadores não anulou os efeitos dos tratamentos clareadores no ES, assim como não teve capacidade de remineralizar o LC.

Characterization of biomodified dentin matrices for potential preventive and reparative therapies.

Biomodification of existing hard tissue structures, specifically tooth dentin, is an innovative approach proposed to improve the biomechanical and biochemical properties of tissue for potential preventive or reparative therapies. The objectives of the study were to systematically characterize dentin matrices biomodified by proanthocyanidin-rich grape seed extract (GSE) and glutaraldehyde (GD). Changes to the biochemistry and biomechanical properties were assessed by several assays to investigate the degree of interaction, biodegradation rates, proteoglycan interaction, and effect of collagen fibril orientation and environmental conditions on the tensile properties. The highest degree of agent-dentin interaction was observed with GSE, which exhibited the highest denaturation temperature, regardless of the agent concentration. Biodegradation rates decreased remarkably following biomodification of dentin matrices after 24h collagenase digestion. A significant decrease in the proteoglycan content of GSE-treated samples was observed using a micro-assay for glycosaminoglycans and histological electron microscopy, while no changes were observed for GD and the control. The tensile strength properties of GD-biomodified dentin matrices were affected by dentin tubule orientation, most likely due to the orientation of the collagen fibrils. Higher and/or increased stability of the tensile properties of GD- and GSE-treated samples were observed following exposure to collagenase and 8 months water storage. Biomodification of dentin matrices using chemical agents not only affects the collagen biochemistry, but also involves interaction with proteoglycans. Tissue biomodifiers interact differently with dentin matrices and may provide the tissue with enhanced preventive and restorative/reparative abilities.

The effect of dentin pretreatment on the microtensile bond strength of self-adhesive resin cements.

STATEMENT OF PROBLEM: Self-adhesive cements have lower bond strength than conventional resin cements that rely on the application of etch-and-rinse adhesive systems. PURPOSE: The purpose of this study was to evaluate the effect of tannic and polyacrylic acid on the microtensile bond strength of self-adhesive resin cements to dentin. MATERIAL AND METHODS: Eighteen molar crowns were flattened to expose dentin and divided into 3 groups according to the dentin pretreatment: (1) control, without surface treatment; (2) 25% polyacrylic acid solution (Ketac Conditioner); (3) 20% tannic acid. Composite resin (Tescera) blocks were luted to dentin surfaces using 2 self-adhesive resin cements (RelyX Unicem or Maxcem Elite) (n=3). All specimens were stored in distilled water for 24 hours, sectioned into beams (1 mm(2)), and tested to failure using a microtensile method at a crosshead speed of 1 mm/min. The data were statistically analyzed using 2-way ANOVA and Fisher's PLSD tests (α=.05). RESULTS: The polyacrylic acid significantly improved the microtensile bond strength of RelyX Unicem (14.92 ±4.94 MPa) when compared to the control group (8.35 ±1.99 MPa) and tannic acid treatment (8.38 ±2.67 MPa) (P<.001). Dentin surface treatment did not affect the microtensile bond strength of Maxcem Elite cemented groups (control, 8.45 ±3.21 MPa; polyacrylic acid, 9.53 ±9.95 MPa; tannic acid, 6.89 ±4.45 MPa). CONCLUSIONS: Dentin pretreatment with polyacrylic acid improved the microtensile bond strength of RelyX Unicem.

Long-term effect of carbodiimide on dentin matrix and resin-dentin bonds.

OBJECTIVES: To characterize the interaction of 1-Ethyl-3-[3-dimethylaminopropyl] carbodiimide Hydrochloride (EDC) with dentin matrix and its effect on the resin-dentin bond. METHODS: Changes to the stiffness of demineralized dentin fragments treated with EDC/N-hydroxysuccinimide (NHS) in different solutions were evaluated at different time points. The resistance against enzymatic degradation was indirectly evaluated by ultimate tensile strength (UTS) test of demineralized dentin treated or not with EDC/NHS and subjected to collagenase digestion. Short- and long-term evaluations of the strength of resin-dentin interfaces treated with EDC/NHS for 1 h were performed using microtensile bond strength (microTBS) test. All data (MPa) were individually analyzed using ANOVA and Tukey HSD tests (alpha = 0.05). RESULTS: The different exposure times significantly increased the stiffness of dentin (p < 0.0001, control-5.15 and EDC/NHS-29.50), while no differences were observed among the different solutions of EDC/NHS (p = 0.063). Collagenase challenge did not affect the UTS values of EDC/NHS group (6.08) (p > 0.05), while complete degradation was observed for the control group (p = 0.0008, control-20.84 and EDC/NHS-43.15). EDC/NHS treatment did not significantly increase resin-dentin muTBS, but the values remained stable after 12 months water storage (p < 0.05). CONCLUSIONS: Biomimetic use of EDC/NHS to induce exogenous collagen cross-links resulted in increased mechanical properties and stability of dentin matrix and dentin-resin interfaces.

The effect of load cycling on nanoleakage of deproteinized resin/dentin interfaces as a function of time.

OBJECTIVES: To evaluate the effects of cyclic loading on nanoleakage as a function of time at resin-dentin interfaces with and without removal of collagen. METHODS: Bovine flat mid-coronal dentin received one of the following surface treatments: (1) acid-etch or (2) acid etched+5% NaOCl for 2m. The teeth were then bonded with Single Bond (3M ESPE), Scotchbond Multi-Purpose (3M ESPE), One-Step Plus (Bisco) or All-Bond 2 (Bisco) adhesive systems and restored with Z250 composite. Half of the bonded teeth were randomly assigned to receive 200,000 cycles of loading at 50 N. Teeth were sectioned into 1 mm x 1 mm thick slices and stored in distilled water for 24h and 6 months. After water storage, beams were prepared for nanoleakage evaluation and observed under the SEM. Data were statistically analyzed using three-way ANOVA and Fisher's PLSD test (p<0.05). RESULTS: Use of NaOCl did not affect nanoleakage at the interface of all adhesive systems (p>0.05) when compared to their respective controls. After 6 months of immersion in water, except for One Step Plus, collagen-depleted groups and control groups presented similar leakage values. After cyclic loading, the deproteinized group revealed a higher degree of silver nitrate deposits when compared to the control group (p<0.05). All four adhesive systems presented a high degree of silver nitrate deposits after 6 months of water storage. SIGNIFICANCE: The hybrid layer is important as a stress-absorbing layer and it may not represent the weak link for initiation of the nanoleakage phenomenon.

Changes in stiffness of demineralized dentin following application of collagen crosslinkers.

It is thought that increasing the strength of the dentin matrix using crosslinking agents may improve both the strength and the durability of resin-dentin bonds. The purpose of this study was to evaluate the effect of two collagen crosslinking agents (glutaraldehyde, GD and grape seed extract, GSE) on the modulus of elasticity of demineralized dentin. Sound molar fragments were fully demineralized and divided into five groups according to the type and concentration of crosslinking agents: 2.5% GD; 5% GD, 25% GD; 0.65% GSE; 6.5% GSE. Specimens were immersed in their respective solution and tested at baseline, 10 min, 30 min, 1 h, 2 h, 4 h. The elastic modulus of dentin was significantly affected by the treatment (p < 0.01) and exposure time (p < 0.01). There was a statistically significant interaction between the two factors evaluated (treatment vs. time p < 0.01). Mean baselines values varied between 4.8 and 6.2 MPa in water; after 4 h of treatment the values increased between 34.9 and 242.5 MPa, that were treatment time and agent dependent. The use of these collagen crosslinkers to increase the stiffness of demineralized dentin, was both concentration and time dependent.

Removal of dentin matrix proteoglycans by trypsin digestion and its effect on dentin bonding.

The aim of this study was to investigate the effect of trypsin digestion on removal of chondroitin sulfate proteoglycans (CS-PGs) in demineralized dentin, and subsequent dentin bonding. Bovine dentin fragments were demineralized, treated with or without trypsin, stained with cupromeronic blue, and observed under transmission electron microscopy. Demineralized sections with or without trypsin digestion were also subjected to immunohistochemical analysis with anti-chondroitin-4-sulfate (C4S) monoclonal antibody, 2-B-6. The presence of galactosamine and glucosamine in the trypsin digest was confirmed by amino acid analysis. Bond strength testing was performed on trypsin treated and control specimens where samples were either kept moist or dried and re-wet, then bonded. Bond strength significantly decreased after trypsin treatment (p < 0.05). TEM, immunohistochemical, and amino acid analyses demonstrated that trypsin digestion efficiently removed C4S-PGs from demineralized dentin matrix. This study indicates that the detrimental effects observed on dentin bonding by trypsinization may be due in part to the removal/cleavage of the C4S-PGs, and further underscore the importance of C4S-PGs on dentin bonding.

Effect of long-term water storage on etch-and-rinse and self-etching resin-dentin bond strengths.

This study sought to evaluate the long-term bond strength of etch-and-rinse and self-etch adhesive systems to dentin after one year of water storage. Crown fragments from the buccal surface of extracted bovine incisors were ground flat to expose dentin surfaces. Four etch-and-rinse and two self-etch bonding agents were used according to manufacturers' instructions. Bonded specimens were stored in water for either 24 hours or one year at 37 delete C. After elapsed storage times, specimens were tested for shear bond strength (SBS) at 0.5 mm/min. Data were analyzed by ANOVA and Tukey's tests (p < 0.05). Both evaluated factors (adhesive system and storage time) and their interactions were statistically significant. Single Bond produced the highest SBS regardless of the water degradation period. One-year water storage reduced the SBS of Prime & Bond NT and One-Up Bond F significantly; the other adhesives performed similarly. Long-term bond strengths of etch-and-rinse and self-etch adhesive systems are susceptible to hygroscopic and hydrolytic effects to varying extents, depending on their chemistry and structure. The presence of monomers with different properties might induce preferential degradation of specific adhesive polymer systems.

Importance of the hybrid layer on the bond strength of restorations subjected to cyclic loading.

The present study evaluated the long-term microtensile bond strength of collagen-depleted dentin followed by cyclic loading. Bovine flat, midcoronal dentin received one of the following surface treatments: acid-etch or acid-etched + 5% NaOCl for 2 min. The teeth were restored with Single Bond, Scotchbond Multi-purpose, One-Step Plus, or All-Bond 2 adhesive systems. Half of the specimens were randomly assigned to receive 200,000 cycles (50 N force). Teeth were sectioned into 1 x 1 mm thick slices and stored for 24 h, 1 month, 3 months, or 6 months. Specimens were subjected to tensile testing after elapsed storage time. Samples were analyzed by three-way ANOVA and Fisher's PLSD (p < 0.05). The results showed that the deproteinized groups without cyclic loading presented lower bond strength when compared with the control group, but the difference was only statistically significant for Single Bond and All-Bond 2 (p < 0.05). All adhesives presented a decrease in bond strength over time, regardless of the dentin treatment. When cyclic loaded, the decrease in bond strength for the deproteinized group was even greater when compared with the control group (p < 0.05). The results suggest that the hybrid layer is important as a stress-buffering layer when loading is applied, and thus the presence of collagen is essential.

Application of crosslinkers to dentin collagen enhances the ultimate tensile strength.

The stabilization of dentin collagen with biocompatible crosslinking agents may be of clinical importance to improve dentin bond strength. The present study aimed to evaluate the effect of three collagen crosslinking agents on the ultimate tensile strength (UTS) of undemineralized and demineralized dentin. Ten freshly extracted sound molars were sectioned into 0.5 x 0.5 mm2 thick beams. The beams were either demineralized or kept undemineralized. Then, specimens were subdivided into four groups according to treatments--PBS solution (control), 5% glutaraldehyde (GD), 0.5% proanthocyanidin PBS solution (PA), and 0.625% genipin PBS solution (GE). Specimens were kept in their respective solutions for either 4 or 40 h. To assess UTS, specimens were subjected to tensile forces at a crosshead speed of 1 mm/min. Statistical analysis was performed using two-way ANOVA and Fisher's PLSD test (p < 0.05). Statistically significant increases in UTS were observed for demineralized dentin after PA and GE dentin treatment, when compared with those of the control group. Dentin treated with GD showed no statistically significant differences in UTS when compared with that the control. Undemineralized dentin revealed no significant differences as compared to that of the control, regardless of the collagen crosslinkers. The application of two naturally occurring crosslinkers, i.e., PA and GE, to dentin collagen significantly improves UTS, indicating its potential value in restorative dentistry.

Microleakage evaluation of a new low-shrinkage composite restorative material.

PURPOSE: This study compared the microleakage of an experimental low-shrinkage resin composite (Hermes), a nanofilled resin composite material (Filtek Supreme) and a hybrid resin composite (Tetric Ceram) using a dye penetration method. METHODS AND MATERIALS: Class I cavities prepared in 60 human molars were randomly divided into 3 groups according to the restorative material used. The preparations were restored using a bulk or an incremental technique. Half of the specimens from each group were subjected to 200,000 cycles of loading at 50 N, while the other half were stored in distilled water for 24 hours at 37 degrees C. All specimens were immersed in 1% methylene blue (pH = 7.0) for 24 hours and sectioned into 3 slabs. The margins were evaluated for microleakage using an ordinal scoring system (0-4) under a stereomicroscope at 40x magnification. Data were subjected to the non-parametric Kruskal-Wallis analysis of variance and Mann-Whitney test (p<0.05). Data were expressed as median leakage scores and mean ranks. RESULTS: All of the restorative systems had microleakage, regardless of the insertion technique and mechanical load cycling. Incremental placement significantly reduced microleakage as compared to the bulk technique, regardless of the restorative system used. Load cycling significantly affected incrementally placed restorations, except for the Hermes system.

Effect of 10% carbamide peroxide dental bleaching on microhardness of filled and unfilled sealant materials.

PURPOSE: The purpose of this study was to quantitatively evaluate the effect of 10% carbamide peroxide on the microhardness of pit and fissure sealant materials. METHODS: Fluroshield, Vitroseal Alfa, and one unfilled (Clinpro) sealants were placed in Teflon matrices (4 mm in diameter by 2 mm in height) and polymerized for 40 seconds. A total of 20 specimens were prepared for each material, in which half were assigned as the control group (stored in artificial saliva and no bleaching treatment). For the remaining half, Clarigel Gold bleaching agent (10% carbamide peroxide) was placed over the specimen surface for 4 hours/day during 4 weeks. When specimens were not under bleaching treatment, they were kept in artificial saliva. Afterwards, specimens were subjected to Knoop microhardness testing using a 25-g load for 5 seconds. Five measurements were made on the sealants' surfaces and then calculated in Knoop hardness values. The data were statistically analyzed by two-way analysis of variance and Tukey's tests with a 5% confidence level. RESULTS: The results of this in vitro study showed that the application of a carbamide peroxide-based bleaching material significantly affected the microhardness values of filled sealant materials. The bleaching agent did not affect the microhardness of the unfilled sealant. CLINICAL SIGNIFICANCE The results of this in vitro study suggest that the bleaching agents altered the surface hardness of filled sealant restorative materials. This could possibly lead to increased wear and surface roughness.

In vitro evaluation of caries inhibition promoted by self-etching adhesive systems containing antibacterial agents.

This study evaluated the cariostatic effect of antibacterial self-etching adhesive systems, by means of an in vitro bacterial caries model. Seventy-five prepared bovine slabs were randomly divided into groups (n=15): (1) unbonded composite, no carious challenge (UNB-NC); (2) unbonded composite, carious challenge (UNB-C); (3) Clearfil SE Bond, no antibacterial agent (CSE); (4) Protect Bond, containing MDPB and fluoride (PB); and (5) Reactmer Bond, fluoride-releasing (RB). All preparations were restored with Filtek Z-250. Groups (2)-(5) were submitted to a medium containing Streptococcus mutans (ATCC-- 25175) for 5 days, and Group (1) was kept in a noninoculated medium. Insoluble polysaccharides present in tooth biofilms were quantified, Knoop hardness (KHN) was measured on the enamel adjacent to restorations, and standard 35-mm polarized light photomicrographs were taken as illustrations. Polysaccharide and Knoop hardness results were analyzed with the use of ANOVA, with a split-split-plot statistical design for KHN. Except for Group (1), all groups showed similar caries formation. Biofilm over PB restorations showed the smallest amounts of polysaccharides (14.37 microg/mg), and CSE showed the highest amounts (20.87 microg/mg). All self-etching systems tested were unable to inhibit secondary caries in a bacterial model simulating a high caries challenge, even though there was reduced glucan synthesis provided by the adhesive system containing MDPB and fluoride.